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Hypoglycemic and antioxidant activities of

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DOI: 10.1016/j.apjtb.2015.03.004

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Original article http://dx.doi.org/10.1016/j.apjtb.2015.03.004

Hypoglycemic and antioxidant activities of Caesalpinia ferrea Martius leaf extract in

streptozotocin-induced diabetic rats
1* 1 1
Sherien Kamal Hassan , Nermin Mohammed El-Sammad , Amria Mamdouh Mousa ,
1 2 3
Maha Hashim Mohammed , Abd el Razik Hussein Farrag , Amani Nassir Eldin Hashim , Victoria
4 4 3
Werner , Ulrike Lindequist , Mahmoud Abd El-Moein Nawwar
1
Department of Biochemistry, National Research Centre, Cairo, Egypt
2
Department of Pathology, National Research Centre, Cairo, Egypt
3
Department of Phytochemistry and Plant Systematics, National Research Centre, Cairo, Egypt
4
Institute of Pharmacy, Pharmaceutical Biology, Ernst-Moritz-Arndt-Universitat¨ Greifswald, D-17487 Greifswald, Germany

ARTICLEINFO ABSTRACT

Article history: Objective: To evaluate the antidiabetic and antioxidant effects of aqueous ethanolic
Received 16 Dec 2014 extract of Caesalpinia ferrea (C. ferrea) leaf in normal and streptozotocin (STZ)
Received in revised form induced diabetic rats.
Received in revised form 31 Dec Methods: Male Sprague-Dawley rats divided into 6 groups of 6 rats each were
2014, 2nd revised form 2 Feb assigned into diabetic and non-diabetic groups. Diabetes was induced in rats by
2015 Accepted 3 Mar 2015 single intraperi-toneal administration of STZ (65 mg/kg body weight). C. ferrea
Available online xxx extract at the doses of 250 and 500 mg/kg body weight was orally administered to
both diabetic and non-diabetic animals for a period of 30 days. After completion of
experimental duration serum, liver and pancreas were used for evaluating
Keywords:
biochemical and histopathological changes.
Caesalpinia ferrea
Diabetes Results: Oral administration of C. ferrea leaf extract significantly reduced elevated
Streptozotocin serum glucose, A-amylase, liver function levels and significantly increased serum
Hypoglycemia insulin, total protein and body weight as well as improved lipid profile due to
Antioxidant markers diabetes. Furthermore, the treatment resulted in a marked increase in glutathione
Histopathology peroxidase, su-peroxide dismutase, catalase and reduced glutathione, and
diminished levels of lipid peroxidation in liver and pancreas of diabetic rats.
Histopathological studies demonstrated the reduction in the pancreas and liver
damage and confirmed the biochemical findings. Conclusions: From the present
study, it can be concluded that the C. ferrea leaf extract effectively improved
hyperglycaemia while inhibiting the progression of oxidative stress in STZ-induced
diabetic rats. Hence, it can be used in the management of diabetes mellitus.

1. Introduction secretion with or without concurrent impairment of insulin ac-

Diabetes mellitus (DM) is a group of metabolic diseases
characterized by abnormal metabolism of carbohydrates, pro-
teins, and fats resulting from inadequate pancreatic insulin
*Corresponding author: Dr. Sherien Kamal Hassan, Biochemistry
Department, National Research Centre, Cairo, Egypt.
Fax: +20 33370931
E-mail: sherien.kamal.hassan@gmail.com
Peer review under responsibility of Hainan Medical University.
Foundation Project: Supported and financed by the Alexander von
Humboldt Foundation through the group linkage programme (joint project:
“Bioactive phenolics from Egyptian folk medicinal plants”, 3.4- Fokoop-
DEU/1093980) awarded to U. L. and M. N.
tion [1]. According to the American diabetes association, the
chronic hyperglycemia is associated with long-term damage,
dysfunction, and failure of different organs, especially the eyes,
kidneys, nerves, heart and blood vessels [2]. DM is considered
the most prevalent disease in the world affecting 25% of the
population. It afflicts 150 million people and is predicted to
rise to 300 million by 2025 [3]. It is likely to be the fifth leading
cause of death worldwide [4].
Previous studies have demonstrated that DM exhibits
enhanced oxidative stress and highly reactive oxygen species
(ROS) production in pancreatic islets due to persistent and
chronic hyperglycemia, thereby depletes the activity of the

2221-1691/Copyright © 2015 Hainan Medical University. Production and hosting by Elsevier (Singapore) Pte Ltd. This is an open access article under the CC
BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

Please cite this article in press as: Hassan SK, et al., Hypoglycemic and antioxidant activities of Caesalpinia ferrea Martius leaf extract in streptozotocin-induced diabetic rats, Asian Pac J
Trop Biomed (2015), http://dx.doi.org/10.1016/j.apjtb.2015.03.004
2 Sherien Kamal Hassan et al./Asian Pac J Trop Biomed 2015; ▪(▪): 1–10
antioxidative defense system, and thus promotes free
radical generation [5]. Oxygen free radicals have been
suggested to be a contributory factor in complications of
DM [6]. It seems to be an oxidative stress-related disorder
and the antioxidants may be useful in preventing it [7].
Therefore supplementation of therapeutics with antioxidants
may have a chemoprotective role in diabetes [8].
Many plant extracts and their products have been shown
to have significant antioxidant effect in treating many kinds
of diseases [9]. The use of medicinal plants for the treatment
of human diseases has increased considerably worldwide
[10]. Ethnopharmacological evidence has shown that the use
of plants is also helpful in prophylaxis or treatment of
diabetes. Given that, herbal medicine possesses
significant efficacy, low incidence of side effects, low
cost and relative safety [11], while synthetic anti-diabetic
agents can produce serious side effects, as hypoglycemic
coma and disturbances of the liver and kidneys [12].
The little studied genus Caesalpinia contains more than
500 species of worldwide distribution [13]. Previous studies
of species of this genus reported remarkable biological
activities such as antimicrobial (Caesalpinia bonducella) [14],
antidiabetic (C. bonducella) [15], antimalarial (Caesalpinia
pluviosa) [16], and anti-inflammatory (Caesalpinia sappan)
[17]. To date, less than 30 species of this genus have been
studied for their phytoconstituents. The metabolites
described include predominantly flavonoid derivatives,
steroids, triterpenoids, and cassane diterpenes [18].
Caesalpinia ferrea (C. ferrea) Martius (Leguminosae),
popularly known as “pau-ferro” or “juca´”, is a large tree
belonging to the Fabaceae family. It is found mainly in the north
and northeast of Brazil. In folk medicine, the tea of the stem
bark of C. ferrea has been used for the treatment of diabetes. In
view of its ethnomedicinal importance, the Brazilian Ministry of
Health has included this species on the national list of
medicinal plants important to the health system [19].
The pharmacological properties of C. ferrea fruits or stem
barks include antiulcerogenic [20], anti-inflammatory [21],
analgesic [22], antibacterial [23], antihypertensive [24],
antidiabetic [19], and cancer chemopreventive [25] activities.
Recently a unique chalcone trimer (pauferrol A) and two
chalcone dimers (pauferrol B and pauferrol C), were isolated
from the stems of C. ferrea. These chalcones exhibited potent
inhibitory activities against topoisomerase II [26,27]. The leaves
contain three formerly unknown di-O-glycosyl-C-glucosyl fla-
vones which were isolated, purified and identified namely:
00 00 00
Iso-vitexin 2 -O-B-[xylopyranosyl-(1
00
xylopyranosyl]; Vitexin 2 -O-B-[xylopyranosyl-(1
00
O-B-xylopyranosyl]; Orientin 2 -O-B-[xylopyranosyl-(1
00
2 )-O-B- xylopyranosyl [28]. However, there is no experimental
evidence proving biological activities of C. ferrea leaf up till now.
Therefore, the present study was aimed to evaluate the possible
hypoglycemic properties of C. ferrea leaf in streptozotocin (STZ)
induced diabetic rats.
2. Materials and methods
2.1. Chemicals
STZ, reduced glutathione, 5,5’-dithiobis (2-nitrobenzoic
acid) (DTNB), 1-chloro 2,4-dinitrobenzene (CDNB) and 2,2-
diphenyl-1-picrylhydrazyl (DPPH) were purchased from
Please cite this article in press as: Hassan SK, et al., Hypoglycemic and antioxidant activities of Caesalpinia ferrea Martius leaf extract in streptozotocin-induced diabetic rats, Asian Pac J
Trop Biomed (2015), http://dx.doi.org/10.1016/j.apjtb.2015.03.004
Sigma–Aldrich (St. Louis, MO, USA). Metaphosphoric acid
(MPA) and nitroblue tetrazolium were purchased from Fluka
(Switzerland), and pyrogallol from Merck (Germany). All
chemicals were of analytical grade.
2.2. Plant material
Leaves of C. ferrea were collected from a tree cultivated
in the Zoological Garden, Cairo, Egypt, in May 2012. The
plant was identified by Prof. Salwa Quashti, National
Research Centre (NRC), Cairo, Egypt. A voucher specimen
(C253) has been deposited at the herbarium of the NRC.
2.3. Plant extraction and isolation
Leaves (2.5 kg), dried in the shadow, were crushed and
exhaustively extracted with 70% (v/v) aqueous EtOH under
reflux (three times, each extraction for 8 h with 2 L). The
ob-tained eluent was dried under vacuum at 55–60 C to
give 200 g aqueous ethanolic extract that was used in the
present study.
2.4. Phytochemical screening
This aqueous ethanol extract of C. ferrea was screened
for the presence of various phytoconstituents such as
steroids, al-kaloids, glycosides, flavonoids, carbohydrates,
amino acids, sa-ponins, terpenoids, tannins, and phenolic
compounds as described by Dawang & Datup and Mythili &
Ravindhran [29,30].
2.5. Determination of the scavenging of DPPH radical
The quantitative DPPH assay was carried out according
to the method of Kedare and Singh [31]. The extract was
dissolved in a concentration of 1 mg/mL in ethanol. From
this stock solution, concentrations of regular dilution were
prepared. Then 500 ML of sample, 375 ML ethanol and
125 ML of 1 mmol/L prepared DPPH solution were placed
together. The test was performed in triplicate. All samples were
incubated in sequence for 30 min in the dark at room
temperature and their absorbance was measured at a
wavelength of 517 nm on UV–vis spectrophotometer
(Shimadzu, Duisburg, Germany). Ascorbic acid was used as
positive control. Percentage of radical scavenging activity
00 (RSA) was calculated as follows:
/2 )-O-B-
00 00 00
/2 )-
RSA% = [(Abs of control – Abs of sample)/Abs of blank] × 100
00 00
/
2.6. Acute toxicity study
The mean lethal dose (LD 50) of the aqueous ethanolic
extract of C. ferrea leaf was determined in rats (weighing 180–
200 g) using the method described by Chinedu et al. [32].
2.7. Experimental animals
Male Sprague-Dawley rats weighting 180–200 g were pur-
chased from the Animal House of National Research Centre,
Egypt. Animals were acclimated for a period of 7 days in our
 Sherien Kamal Hassan et al./Asian Pac J Trop Biomed 2015; ▪(▪): 1–10
laboratory condition prior to the experiment. The animals were
fed with standard laboratory diet and allowed to drink water ad
libitum under well ventilated conditions of 12 h light/dark cy-
cles. Experimental protocols for the animal studies were carried
out in accordance with Institutional Ethical Guidelines for the
care of laboratory animals of the National Research Centre.
2.8. Induction of diabetes
Diabetes was induced in overnight fasted rats by a single
intraperitoneal (i.p.) injection of a freshly buffered (0.1 mol/L
citrate, pH 4.5) solution of STZ at a dosage of 65 mg/kg body
weight. After 72 h of STZ administration, the tail vein blood was
collected to determine fasting blood glucose level with an Accu-
Chek sensor comfort glucometer (China). Only rats with hy-
perglycemia (glucose over 250 mg/dL) were considered
diabetic and included in the experiment.
2.9. Experimental design
Rats were randomly divided into six groups, comprising
six rats each. The treatment schedule was as follows:
Group I: normal control (NC); Group II: C. ferrea leaf extract
(500 mg/kg body weight) treated normal rats (CF500-NC);
Group III: C. ferrea leaf extract (250 mg/kg body weight) treated
normal rats (CF250-NC); Group IV: diabetic control (DC);
Group V: C. ferrea leaf extract (500 mg/kg body weight) treated
diabetic rats (CF500-DC); Group VI: C. ferrea leaf extract (250
mg/kg body weight) treated diabetic rats (CF250-DC).
Different doses of C. ferrea aqueous ethanolic extract
were administered orally using an intragastric tube daily to
the respective group till the end of the experiment. All doses
were started 72 h after STZ injection.
2.10. Blood and tissue sampling
At the end of the 30-day experiment (after diabetes induction),
overnight fasting animals were ether anaesthetized. Venous retro
orbital blood samples were collected using a glass capillary without
anticoagulant. Serum was separated by centrifugation at 3000
r/min for 15 min. The resulting samples were stored at −20 C until
assayed. Liver and pancreas were removed and washed in ice-
cold saline solution immediately, and then each organ was divided
into two portions. A portion was homogenized in 0.1 mol/L
potassium phosphate buffer (pH 7.4) using Tissue master TM125
(Omni International, USA). After centrifugation at 3000 r/min for 10
min, the clear supernatant was stored at −80 C to be used for
biochemical analysis. The other portion of the liver and pancreas
was fixed in 10% formalin for histological analysis.
2.11. Biochemical analysis
2.11.1. Determination of serum glucose, insulin and A-
amylase
Blood glucose was determined using Biodiagnostic kit,
Egypt. Insulin level was estimated with sandwich immunolu-
minometric assay kit supplied from Snibe Co., Ltd., China
using Maglumi 1000 fully automated chemiluminescence
immuno-assay analyzer (Snibe Co.,Ltd., China). Alpha-
amylase activity was assayed using kits supplied by ELitech
Clinical Systems, France.
Please cite this article in press as: Hassan SK, et al., Hypoglycemic and antioxidant activities of Caesalpinia ferrea Martius leaf extract in streptozotocin-induced diabetic rats, Asian Pac J
Trop Biomed (2015), http://dx.doi.org/10.1016/j.apjtb.2015.03.004
3
2.11.2. Determination of serum lipid profile
Serum concentrations of triglyceride (TG), total cholesterol
(TC), and high-density lipoprotein cholesterol (HDL-C) were
determined using commercially available kits supplied by
Reactivos GPL, Spain. Low-density lipoprotein cholesterol
(LDL-C) was calculated according to Friedewald's formula
[33]:
LDL = [(TC − HDL) − TG/5]
2.11.3. Determination of serum liver function
Serum aspartate transaminase (AST) and alanine trans-
aminase (ALT) were assayed using kits provided by Biorexfars,
UK. Serum alkaline phosphatase (ALP) was estimated using
kits supplied by Stanbio, USA, whereas serum glutamyl trans-
peptidase (GGT) and serum total protein (TP) were measured
using kits supplied by Reactivos GPL, Spain and Biodiagnostic,
Egypt respectively.
2.11.4. Determination of oxidative stress markers in
hepatic and pancreatic tissue
Glutathione peroxidase activity (GSH-Px) was measured
according to the method of Necheles et al. [34]. Superoxide
dismutase activity (SOD) was investigated utilizing the
technique of Minami and Yoshikawa [35]. Catalase (CAT)
activity was determined by the method of Aebi [36]. Lipid
peroxidation was estimated colorimetrically by measuring
thiobarbituric acid reactive substances (TBARS) according
to method of Lefevre` et al. [37]. Reduced glutathione (GSH)
was estimated according to the method of Beutler et al. [38]
after precipitating liver and pancreas proteins with 10%
MPA.
2.11.5. Histopathological investigation
The histopathologic examination was performed by light
microscopy on liver and pancreas specimens that were fixed
in 10% formalin. After fixation, the samples were processed to
obtain 5 Mm thick paraffin sections. Pancreas and liver
sections were stained with hematoxilin and eosin (H & E). Then
the slides were observed under a Leica photomicroscope.
2.11.6. Image morphometry
The morphometric analysis was performed at the Pathol-
ogy Department, National Research Center using the Leica
Qwin 500 Image Analyzer (LEICA Imaging Systems Ltd.,
Cambridge, England) which consists of Leica DM-LB mi-
croscope with JVC color video camera attached to a com-
puter system Leica Q 500IW. The morphometric analysis is
carried out on H & E stained slides. The slides to be
examined were placed on the stage of the microscope, and
focused it at low power magnification (100×). We screen
the slide to determine the boundaries of the tissue to be
measured. The condenser is centered and focused, and the
light source is set to the required level. Successful
adjustment of illumination is checked for on the video
monitor. The area of Langerhans islets were measured by
drawing a line starting from one edge to the other and from
one edge till the opposite, respectively. The results appear
automatically on the monitor in the form of square micron
2
(Mm ) with the mean and standard error.
4 Sherien Kamal Hassan et al./Asian Pac J Trop Biomed 2015; ▪(▪): 1–10

2.12. Statistical analysis 3.4. Effect of C. ferrea extract on body weight

Data were expressed as mean ± SEM. The statistical signif- As shown in Table 2, body weights of rats in DC group were
icance was evaluated by One-way analysis of variance lower than those in other groups. STZ caused a significant
(ANOVA) using SPSS-14 statistical software followed by LSD weight loss of rats in DC and CF250-DC groups in comparing
test to detect differences between groups. The differences were to NC group, while treatment with C. ferrea extract at 500 mg/kg
considered statistically significant at P < 0.05. body weight to diabetic rats significantly suppressed such
decrease in the body weight. No significant difference was
3. Results observed after treatment with C. ferrea extract in CF250-DC as
compared to the DC group.
3.1. Phytochemical screening of C. ferrea extract
Table 2
The preliminary phytochemical screening of C. ferrea Changes of the body weight of rats during the experimental period
aqueous ethanolic extract indicated the presence of of 30 days.
carbohy-drates, glycosides, tannins, and phenolic Groups Initial body weight (g) Final body weight (g)
compounds as shown in Table 1.
I) NC 185.00 ± 3.41 235.44 ± 6.24
II) CF500-NC 181.33 ± 1.74 230.92 ± 7.19b
III) CF250-NC 198.33 ± 4.01 235.50 ± 10.17b
Table 1
IV) DC 190.00 ± 4.47 182.33 ± 5.67a
Phytochemical screening of aqueous ethanolic extract of C. ferrea leaf. V) CF500-DC 183.66 ± 2.07 218.00 ± 4.53b
Phytochemicals Presence/absence VI) CF250-DC 189.83 ± 3.74 198.70 ± 6.68a
Carbohydrates and/or glycosides Present Data are expressed as mean ± SEM (n = 6). Values with different su-
Tannins Present perscripts down the column are significantly different at P < 0.05.
Saponins Absent a
Statistically different from NC group.
b
Alkaloids Absent Statistically different from DC group.
Anthraquinones Absent
Unsaturated sterols or triterpenes Absent
Phenolic compounds Present 3.5. Effect of C. ferrea extract on blood glucose, insulin
and A-amylase
3.2. Radical scavenging activity of C. ferrea extract
As shown in Table 3, serum glucose levels of DC and CF-DC
groups were significantly increased as compared to NC group.
During evaluation of the antioxidant activity, the aqueous
ethanol extract of C. ferrea exhibited a remarkable radical The administration of C. ferrea extract to STZ-induced diabetic rats
scavenging activity in the DPPH assay. Figure 1 in groups CF500-DC and CF250-DC significantly reduced serum
demonstrates this effect quantitatively in comparison to glucose levels as compared to the DC group. Whereas, serum
those of ascorbic acid. The antioxidant capacity of the insulin levels of DC and CF250-DC groups significantly
extract (ED50) was determined to be (12.45 ± 2.86) Mg/mL. decreased as compared to NC group. The administration of 500
and 250 mg/kg of C. ferrea extract to diabetic rats significantly
3.3. Acute toxicity increased insulin level as compared to the DC group and nearly
returned to the basal level in a dose-dependent manner. More-over,
A-amylase activity in the DC and CF-DC groups were significantly
Acute toxicity studies revealed the non-toxic nature of C. higher than those in the normal NC group. The administration of C.
ferrea aqueous ethanolic extract as the treated rats appeared ferrea extract to diabetic rats in CF500-DC
normal and did not display any significant changes in
behavior or neurological responses up to 1500 mg/kg body
weight of the extract. There was no mortality or toxicity reaction
at any of the doses until the end of the study. Table 3
Serum glucose, insulin, and A-amylase values in all groups.
Groups Glucose Insulin A-amylase
(mg/dL) (MIU/mL) (IU/L)
I) NC 102.14 ± 3.87 2.80 ± 0.25 15.04 ± 0.68
II) CF500-NC 95.86 ± 2.99b 3.07 ± 0.20b 16.75 ± 0.97b
III) CF250-NC 96.84 ± 2.66b 2.92 ± 0.18b 14.85 ± 0.66b
IV) DC 388.49 ± 19.00a 1.23 ± 0.05a 249.50 ± 15.83a
V) CF500-DC 121.60 ± 5.32a,b 2.47 ± 0.13b 153.83 ± 4.98a,b
Change from 68.60% 101.6% 38.34%
DC (%)
VI) CF250-DC 134.50 ± 8.76a,b 2.02 ± 0.10a,b 174.71 ± 6.63a,b
Change from 65.37% 64.48% 29.97%
DC (%)
Data are expressed as mean ± SEM (n = 6). Values with different su-
perscripts down the column are significantly different at P < 0.05.
a b
Figure 1. Antioxidant capacity of the aqueous ethanolic extract of C. Statistically different from NC group.
ferrea leaf (DPPH assay). Statistically different from DC group.

Please cite this article in press as: Hassan SK, et al., Hypoglycemic and antioxidant activities of Caesalpinia ferrea Martius leaf extract in streptozotocin-induced diabetic rats, Asian Pac J
Trop Biomed (2015), http://dx.doi.org/10.1016/j.apjtb.2015.03.004
 Sherien Kamal Hassan et al./Asian Pac J Trop Biomed 2015; ▪(▪): 1–10
5

Table 4
Effect of C. ferrea extract on serum lipid profile in all groups.

Groups TG (mg/dL) TC (mg/dL) HDL-C (mg/dL) LDL-C (mg/dL)

I) NC 48.39 ± 1.89 52.05 ± 1.57 28.71 ± 0.80 13.77 ± 1.23
II) CF500– NC 48.14 ± 1.93b 47.47 ± 1.43b 27.54 ± 0.96b 10.28 ± 0.98b
III) CF250– NC 28.46 ± 1.92a,b 46.74 ± 1.37b 29.62 ± 0.97b 11.43 ± 1.04b
IV) DC 108.17 ± 6.37a 99.59 ± 5.44a 18.36 ± 0.63a 59.60 ± 5.23a
V) CF500- DC 49.35 ± 1.36b 58.91 ± 2.63b 25.20 ± 1.40a,b 22.27 ± 2.28a,b
Change from DC (%) 54.37% 40.84% 37.25% 62.63%
VI) CF250- DC 61.14 ± 2.82a,b 56.38 ± 1.46b 24.41 ± 1.10a,b 21.32 ± 1.37a,b
Change from DC (%) 43.47% 43.38% 32.95% 64.22%
Data are expressed as mean ± SEM (n = 6). Values with different superscripts down the column are significantly different at P < 0.05.
a b
Statistically different from NC group.
Statistically different from DC group.

and CF250-DC groups significantly decreased A-amylase ac- group as compared to normal control rats. Administration of 500
tivity as compared to the DC group. and 250 mg/kg of C. ferrea extract for diabetic rats significantly
increased TP level and adjusted to the normal level.
3.6. Effect of C. ferrea extract on lipid profile
Table 4 shows the levels of serum lipid profile of rats in
different experimental groups. Rats in DC group displayed a
significant increase in the levels of TG, TC, and LDL-C in
comparison with NC group. However, serum HDL-C level of
rats in DC group was significantly lower than that of rats in
NC group. Treatment with C. ferrea extract in CF500-DC
and CF250-DC groups showed a significant decrease in
the levels of serum TG, TC, and LDL-C and simultaneous
significant in-crease in the level of HDL-C when compared
with DC group. Although the serum HDL-C and LDL-C level
did not return to the basal level of NC group, the serum TG
level in CF500-DC group and TC level in both CF500-DC
and CF250-DC groups were able to return.
3.7. Effect of C. ferrea extract on serum liver function
The data for serum liver function tests are presented in
Table 5. Serum activities of AST, ALT, ALP, and GGT bio-
markers of liver toxicity were significantly elevated in STZ
induced diabetic rats when compared to normal controls.
Treatment of diabetic rats with 500 and 250 mg/kg of C. ferrea
extract significantly reduced the activity of these biomarkers
with respect to diabetic control rats for both doses. Such
reduction nearly returned to the basal normal level for AST and
ALT activities but ALP and GGT could not return. On the
contrary, serum level of TP was significantly decreased in DC
3.8. Effect of C. ferrea extract on hepatic and pancreatic
oxidative stress markers
Table 6 reveals a significant decrease in antioxidant
enzyme activities (GSH-Px, SOD, CAT) and antioxidant GSH
level. A significant elevation in TBARS production was
observed in the hepatic and pancreatic tissues of rats in DC
group when compared with NC group. C. ferrea extract
treatment of diabetic rats in CF500-DC and CF250-DC groups
significantly raised GSH-Px, SOD & CAT enzyme activities
and GSH level and inhibited the formation of TBARS as
compared to DC group in a dose-dependent manner. Though,
such improvement in GSH and TBARS levels did not restore to
basal level of NC group, while GSH-Px, SOD and CAT enzyme
activities returned to normal basal values in CF500-DC.
3.9. Effect of C. ferrea extract on pancreas and liver
histopathological examination
The histological investigation of pancreas showed normal
architecture in case of NC, CF500-NC and CF250-NC groups. The
endocrine portions of pancreas or islets of Langerhans were
present in the pancreatic tissue featured circular shapes with
normal cell lining, while the exocrine components that included
acini appeared well organized and with normal morphology. The
interlobular duct was surrounded with the supporting tissue (Figure
2A–C). The image analyzer results showed that the

Table 5
Effect of C. ferrea extract on the activity of liver enzymes and total protein in all groups.
Groups AST (IU/L) ALT (IU/L) ALP (IU/L) GGT (IU/L) TP (g/dL)
I) NC 44.00 ± 2.61 44.67 ± 1.93 23.24 ± 1.52 1.57 ± 0.16 5.55 ± 0.05
II) CF500- NC 45.66 ± 1.52b 43.92 ± 1.20b 25.36 ± 1.54b 1.40 ± 0.18b 5.70 ± 0.10b
III) CF250- NC 46.16 ± 1.82b 49.66 ± 1.91b 26.86 ± 1.17b 1.64 ± 0.05b 5.65 ± 0.18b
IV) DC 80.50 ± 3.33a,b 59.33 ± 1.89a 56.77 ± 2.03a 3.30 ± 0.38a 4.85 ± 0.18a
V) CF500- DC 50.16 ± 3.71b 46.50 ± 1.43b 36.77 ± 2.28a,b 2.54 ± 0.22a,b 5.73 ± 0.21b
Change from DC (%) 37.68% 21.62% 35.22% 23.03% 18.14%
VI) CF250- DC 51.66 ± 4.96b 47.83 ± 1.90b 37.63 ± 1.86a,b 2.57 ± 0.21a,b 5.53 ± 0.14b
Change from DC (%) 35.82% 19.38% 33.71% 22.12% 14.02%
Data are expressed as mean ± SEM (n = 6). Values with different superscripts down the column are significantly different at P < 0.05.
b
a Statistically different from NC group.
Statistically different from DC group.

Please cite this article in press as: Hassan SK, et al., Hypoglycemic and antioxidant activities of Caesalpinia ferrea Martius leaf extract in streptozotocin-induced diabetic rats, Asian Pac J
Trop Biomed (2015), http://dx.doi.org/10.1016/j.apjtb.2015.03.004
6 Sherien Kamal Hassan et al./Asian Pac J Trop Biomed 2015; ▪(▪): 1–10

Table 6
Effect of C. ferrea extract on oxidative stress markers of liver and pancreas in all groups.

Groups GSH-Px (IU/g tissue) SOD (IU/g tissue) CAT (IU/mg tissue) GSH (mg%) TBARS (nmol/mg tissue)
Liver
I) NC 74.11 ± 1.54 99.37 ± 2.53 16.12 ± 0.67 12.09 ± 0.91 14.16 ± 0.92
II) CF500- NC 83.10 ± 1.93b 116.13 ± 1.67b 18.90 ± 0.78b 13.24 ± 0.87b 13.67 ± 0.74b
III) CF250- NC 76.94 ± 1.28b 102.53 ± 5.33b 17.89 ± 0.57b 11.70 ± 1.30b 14.78 ± 0.57b
IV) DC 59.70 ± 2.74a 73.40 ± 3.45a 10.72 ± 0.57a 1.20 ± 0.30a 25.74 ± 0.24a
V) CF500- DC 71.63 ± 1.12b 93.06 ± 2.93b 16.21 ± 0.65b 7.68 ± 2.26a,b 16.81 ± 0.45a,b
Change from DC (%) 19.98% 26.78% 51.21% 540% 34.69%
VI) CF250- DC 67.29 ± 1.13a,b 84.40 ± 2.27a,b 14.37 ± 0.61b 7.11 ± 1.68a,b 19.36 ± 0.28a,b
Change from DC (%) 12.71% 14.98% 34.04% 492.5% 24.78%
Pancreas
I) NC 54.74 ± 1.84 67.28 ± 2.94 2.57 ± 0.17 4.29 ± 0.46 6.22 ± 0.47
II) CF500- NC 62.63 ± 1.87b 71.70 ± 3.19b 3.06 ± 0.22b 4.97 ± 0.47b 5.52 ± 0.54b
III) CF250- NC 58.53 ± 2.25b 64.69 ± 1.97b 2.89 ± 0.25b 4.44 ± 0.23b 4.72 ± 0.38a,b
IV) DC 41.85 ± 2.36a 43.63 ± 2.35a 0.68 ± 0.08a 0.37 ± 0.02a 13.22 ± 0.15a
V) CF500- DC 52.72 ± 1.98b 62.86 ± 2.54b 2.37 ± 0.16b 1.71 ± 0.16a,b 8.67 ± 0.43a,b
Change from DC (%) 25.97% 44.07% 284.52% 362.16% 34.41%
VI) CF250- DC 48.89 ± 2.14b 56.26 ± 1.99a,b 2.06 ± 0.17b 1.32 ± 0.13a,b 9.59 ± 0.32a,b
Change from DC (%) 16.82% 28.94% 202.94% 256.75% 27.45%
Data are expressed as mean ± SEM (n = 6). Values with different superscripts down the column are significantly different at P < 0.05.
a b
Statistically different from NC group.
Statistically different from DC group.

mean islets area in non-diabetic rats was (163.30 ± 5.62)
2
Mm , whereas for CF500-NC and CF250-NC groups were
2 2
(191.23 ± 1.05) and (171.17 ± 2.02) Mm , respectively.
In case of pancreas of diabetic rats, histopathological exam-
ination of pancreas showed the acinar cells around the islets
though seemed to be in normal proportion did not look
classical. The cells of islets were in degenerative form with
asymmetrical vacuoles. Intra islets hemorrhage was also seen
(Figure 2D and E). A significant reduction in the number of B-
cells and size of islet cells was detected. The mean islets area
2
of the diabetic rats was (122.93 ± 15.13) Mm . These indicate it
appeared smaller in comparison with normal rats.
Microscopic investigation of pancreas sections of CF500-DC
and CF250-DC groups revealed regeneration and restoration of
size of Langerhans’ islets along with B–cells repair (Figure 2F and
G), suggesting a protective effect on the islets. This recovery
of the B-cell was more evident at higher dose, whereas the
mean of islets area for CF500-DC and CF250-DC groups were
(196.33 ± 14.55) and (192.12 ± 5.22) Mm , respectively.
The microscopic examinations of sections of liver of NC,
CF500-NC and CF250-NC groups showed the normal structure of
the hepatic lobule. The central vein is surrounded by the
hepatocytes with eosinophilic cytoplasm and distinct nuclei. The
hepatic sinusoids were shown between the hepatocytes (Figure
3A–C). Microscopic examination of liver of DC rats indicated
congestion in the portal tract that was associated with necrosis of
the hepatocytes that surrounded it and moderated inflammatory
infiltration. Some of the nuclei of the hepatocytes revealed
pyknotic form (Figure 3D). In CF500-DC rats, the hepatic lobule
appeared more or less like control. The activated Kupffer cells in
the sinusoids were seen (Figure 3E). In some rats congested portal
tract associated with necrosis of the

Figure 2. The histological investigation of pancreas. A, B, C: Pancreas sections of NC, CF500-NC and CF250-NC groups respectively, showing dense-
staining acinar cells and a light-staining islet of Langerhans just right of the canter of the field; D: Diabetic rat showing the acinar cells around the is-
lets though seemed to be in normal proportion did not look classical. The islet is shrunken and associated with intra islet hemorrhage (arrow). E: Diabetic rat
showing degenerative islet of Langerhans (asterisk) associated with different size of vacuoles (long arrow) and hemorrhage (short arrow); F: CF500-DC rat
showing the exocrine pancreas appearing more or less as control. Few degenerative cells are seen in the islet. G: CF250-DC rat showing islet of Langerhans
that appeared relatively larger than the control one. Exocrine pancreas appeared more or less as control (H & E, Scale bar: 20 Mm).

Please cite this article in press as: Hassan SK, et al., Hypoglycemic and antioxidant activities of Caesalpinia ferrea Martius leaf extract in streptozotocin-induced diabetic rats, Asian Pac J
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 Sherien Kamal Hassan et al./Asian Pac J Trop Biomed 2015; ▪(▪): 1–10
7

Figure 3. The microscopic examinations of sections of liver of NC, CF500-NC and CF250-NC groups. A, B, C: Liver sections of NC, CF500-
NC and CF250-NC groups, respectively, showing the normal architecture of a hepatic lobule and hepatocytes; D: Diabetic group showing
congested portal tract that is associated with necrosis of the hepatocytes that surround it and moderate in flammatory infiltration. Some
of the nuclei of the hepatocytes are pyknotic. E: CF500-DC rat showing hepatic lobule that appears more or less like control. Notice the
activated Kupffer cells. F: CF500-DC rat showing congested portal tract that is associated with necrosis of the hepatocytes that surround it
and moderate inflammatory infiltration. G: CF250-DC rat showing the hepatocytes appearing more or less as the control; H: CF250-DC
rat showing mild congestion of the portal tract that is associated with few inflammatory infiltrations (H & E, Scale bar: 20 Mm).

hepatocytes that surrounded it and moderated inflammatory
infiltration were found (Figure 3F). In CF250-DC rats, the liver
examination showed that the hepatocytes appeared more or
less as the control (Figure 3G), while in some cases mild
congestion of the portal tract associated with few
inflammatory infiltration was seen (Figure 3H).
4. Discussion
Diabetes mellitus is currently a major public health
concern, because its incidence and prevalence are
elevated and increasing, reaching epidemic proportions [39].
Cumulative evidence has shown that poorly and erratically
controlled hyperglycemia produces abnormally high levels
of ROS [40], and these reactive substances could react with
essential molecules such as lipids, proteins and DNA,
leading to histological changes as well as functional
alterations [41]. STZ is a toxin frequently used to induce
diabetes in experimental animals through its ability to
induce selective destruction of pancreatic beta cells
resulting in insulin deficiency and hyperglycemia [42]. To
the best of our knowledge, this is the first report that
analyzes hypoglycaemic effect of C. ferrea leaf aqueous
ethanolic extract on STZ induced experimental diabetes.
In the present study, reduction in body weight in diabetic
rats was observed which might be the result of degradation
of structural proteins due to unavailability of carbohydrates
for utilization as an energy source [43,44]. These results
agree with previous observations that have also reported
loss of body weight [45,46]. A significant increase was
observed in body weight of diabetic rats treated with C.
ferrea extract (CF500-DC group) as compared to diabetic
group which indicates the preventive effect of the extract on
degradation of structural proteins.
The diabetic rats were found to have higher glucose level and
lower level of insulin when compared to normal control rats. From
the results of the present experiment, it was observed that
treatment with C. ferrea extract decreased the serum glucose and
increased serum insulin in STZ induced diabetic rats (CF-DC
groups). It is perhaps due to stimulation of insulin secretion from
remnant pancreatic B-cells, which in turn enhances glucose
utilization by peripheral tissues of diabetic rats either by pro-moting
glucose uptake and metabolism, or by inhibiting hepatic
gluconeogenesis [47]. This is confirmed by histopathological
observations which show that the structural integrity of islets of
Langerhans was restored towards normalization.
Alpha-amylase is one of the main enzymes in human body
that is responsible for the breakdown of starch to more simple
sugars. A-amylases hydrolyze complex polysaccharides to pro-
duce oligosaccharides and disaccharides which are then hy-
drolyzed by A-glycosidase to monosaccharides which are
absorbed through the small intestines into the hepatic portal
vein and increase postprandial glucose levels [48]. In our
investigation, a significant increase in A-amylase was
observed in diabetic rats as compared to control. This result is
in agreement with Adaramoye [49]. Treatment with C. ferrea
extract in CF500-DC and CF250-DC groups moder-ately
inhibited A-amylase. In our phytochemical screening, we have
proved the presence of phenolic compounds in C. ferrea
extract. Some phenolic compounds are known to inhibit the
activity of carbohydrate hydrolyzing enzymes like A-amylase
and A-glucosidase [50].
In diabetes, hyperglycemia is accompanied with dyslipidemia
representing risk factor for coronary heart diseases. The abnormal
high level of serum lipids is mainly due to the uninhibited actions of
lipolytic hormones on the fat depots, mainly due to the action of
insulin. Under normal circumstances, insulin activates the enzyme
lipoprotein lipase, which hydrolyzes TGs. However, in diabetic state
lipoprotein lipase is not activated due to insulin deficiency,
resulting in hypertriglyceridemia, and insulin defi-ciency is also
associated with hypercholesterolemia due to metabolic
abnormalities [51]. TGs stimulate the secretion of very low-density
lipoprotein cholesterol and such increase in very low-density
lipoprotein cholesterol particles reduces the HDL-C level and
increases the LDL-C particles [52]. The characteristic features of
diabetic dyslipidemia are increase in serum TG, TC,

Please cite this article in press as: Hassan SK, et al., Hypoglycemic and antioxidant activities of Caesalpinia ferrea Martius leaf extract in streptozotocin-induced diabetic rats, Asian Pac J
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8 Sherien Kamal Hassan et al./Asian Pac J Trop Biomed 2015; ▪(▪): 1–10
LDL-C, and fall in HDL-C levels [53]. In our study, the altered
serum lipid profile was found in diabetic rats. This finding is
in correlation with the findings of Pepato et al. and Sharma et
al. [54,55]. This altered serum lipid profile was reversed towards
normal after administration of C. ferrea extract with both doses.
Thus, the extract could be helpful in improving lipid metabolism
which will in turn help to prevent diabetic complications such as
coronary heart diseases and atherosclerosis.
It has been well established that elevated levels of AST, ALT
and ALP are indicative of cellular leakage and loss of functional
integrity of the hepatic cell membranes implying hepatocellular
damage [56]. In the present study, the injection of STZ induces
hepatocellular damage, which is one of the characteristic
changes in diabetes as evidenced by high serum levels of AST,
ALT, ALP and GGT in diabetic group compared to the normal
control, suggesting possible damage to the liver. Liver damage
in diabetic rats was confirmed. However, diabetic groups
treated with C. ferrea extract in CF500-DC and CF250-DC
groups showed a significant reduction in the levels of these
enzymes when compared to the diabetic untreated control,
which consequently alleviated the damage caused by STZ as
confirmed by hepatocytes morphology. This means that C.
ferrea has some hepatoprotective potentials in diabetic rats by
decreasing serum AST, ALT, ALP and GGT levels. Treatment of
normal rats with C. ferrea in CF500-NC and CF250-NC groups
maintained the levels of serum AST, ALT, ALP and GGT thereby
showing its non-toxic nature.
Under condition of severe oxidative stress, free radical gen-
eration leads to protein modification. Proteins may be damaged
directly by specific interactions of free radicals with particular
susceptible amino acids [57]. The finding of our study revealed a
significant decrease in the level of serum TP in diabetic rats. This
could be due to increased peroxidation. On the other hand, C.
ferrea treated rats showed increased level of TP, suggesting that C.
ferrea extract has antioxidant capacity.
Oxidative stress is suggested as mechanism underlying
dia-betes and diabetic complications, which results from an
imbal-ance between radical generating and radical
scavenging systems [6]. Antioxidant enzymes as well as
nonenzymatic antioxidants are first line of defense against
ROS induced oxidative damage in a living organism [58]. SOD,
CAT and GSH-Px are the three major scavenging enzymes that
remove the toxic free radicals in vivo [7]. SOD protects tissues
against oxygen free radicals by catalyzing the removal of
superoxide radical, converting it into H 2O2 and molecular
oxygen, which both damage the cell membrane and other
biological structures. CAT is a haemprotein, which is
responsible for the detoxification of significant amounts of
H2O2 [59]. GSH-Px plays a central role in the catabolism of
H2O2 and the detoxification of endogenous metabolic
peroxides and hydroperoxides, which catalyzes GSH [60].
Glutathione functions as a free radical scavenger and is an
essential co-substrate for GSH-Px [61].
The decreased activity of antioxidant (GSH-Px, SOD and
CAT) enzymes along with decreased GSH level was found in
the liver and pancreatic tissues of diabetic rats. These results
are in agreement with Cheng et al. [62]. It was suggested that
decreased antioxidant enzyme activity in DC group could be
due to glycation of these enzymes, which occurred at
persistently elevated blood glucose levels [63]. However,
administration of C. ferrea extract in CF500-DC, CF250-DC
Please cite this article in press as: Hassan SK, et al., Hypoglycemic and antioxidant activities of Caesalpinia ferrea Martius leaf extract in streptozotocin-induced diabetic rats, Asian Pac J
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groups increased the GSH-Px, SOD and CAT activities and
GSH level in the liver and pancreas of diabetic rats.
Lipid peroxidation is one of the characteristic features of
chronic diabetes. The increased free radicals produced may
react with polyunsaturated fatty acids in cell membranes
leading to lipid peroxidation. Lipid peroxidation will in turn result
in the elevated production of free radicals [64]. In the present
study, it was observed that TBARS level in liver and pancreas
of STZ-induced diabetes was significantly increased when
compared to the control. The decreased activity of antioxidant
molecules along with elevated TBARS level in diabetic rats
could probably be associated with oxidative stress and
decreased antioxidant defense potential [7]. Diabetic rats
treated with C. ferrea extract in CF500-DC, CF250-DC groups
showed decreased level of TBARS.
Robertson et al. demonstrated that antioxidants have been
shown to break the worsening of diabetes by improving B-cells
function in animal models and suggested that enhancing anti-
oxidant defense mechanisms in pancreatic islets may be a
valuable pharmacologic approach to managing diabetes [65].
In the present study, the biochemical findings observed
in diabetic rats are in conformity with histopathological
alterations of B-cells of pancreas and hepatocytes. Such
histopathological alterations were reduced by administration
of C. ferrea extract at both doses.
It can be concluded that the aqueous ethanolic extract of C.
ferrea leaf has potential antihyperglycemic activity in STZ
induced diabetic rats. In this sense, the antidiabetic effect may
be due to the presence of secondary metabolites like phenols
and flavonoids in the C. ferrea leaf extract which are
responsible for antioxidant actions and have been found to be
beneficial in controlling diabetes as evident from earlier
studies. The three new phenolic compounds (isovitexin, vitexin
and orientin de-rivatives) isolated in a previous study showed
high antioxidant properties (results not shown) and may
contribute to the major antioxidant activity of the C. ferrea leaf
extract [28]. Experimental evidence obtained from this study is
encouraging enough to warrant further studies on the leaf
extract of this plant to find out its mechanism of action and to
establish its therapeutic potential in the prophylaxis and/or
treatment of diabetes and diabetic complications.
Conflict of interest statement
We declare that we have no conflict of interest.
Acknowledgments
This research was supported and financed by the
Alexander von Humboldt Foundation through the group
linkage pro-gramme (joint project: “Bioactive phenolics from
Egyptian folk medicinal plants”, 3.4- Fokoop-DEU/1093980)
awarded to U. L. and M. N.
References
[1] Ortiz-Andrade RR, Garc´ıa-Jimenez´ S, Castillo-España P,
Ram´ırez-Avila G, Villalobos-Molina R, Estrada-Soto S. alpha-
Glucosidase inhibitory activity of the methanolic extract from
Tournefortia hartwegiana: an anti-hyperglycemic agent. J
Ethnopharmacol 2007; 109(1): 48-53.
 Sherien Kamal Hassan et al./Asian Pac J Trop Biomed 2015; ▪(▪): 1–10
[2] American Diabetes Association (ADA). Diagnosis and classi fica-
tion of diabetes mellitus. Diabetes Care 2012; 33(Suppl. 1): S62-9.
[3] Biswas M, Kar B, Bhattacharya S, Kumar RB, Ghosh AK,
Haldar PK. Antihyperglycemic activity and antioxidant role of
Terminalia arjuna leaf in streptozotocin-induced diabetic rats.
Pharm Biol 2011; 49(4): 335-40.
[4] Roglic G, Unwin N, Bennett PH, Mathers C, Tuomilehto J, Nag
S, et al. The burden of mortality attributable to diabetes:
realistic estimates for the year 2000. Diabetes Care 2005; 28(9):
2130-5.
[5] Savu O, Ionescu-Tirgoviste C, Atanasiu V, Gaman L, Papacocea
R, Stoian I. Increase in total antioxidant capacity of plasma despite
high levels of oxidative stress in uncomplicated type 2 diabetes
mellitus. J Int Med Res 2012; 40(2): 709-16.
[6] Neethu P, Haseena P, ZevaluKezo, Thomas SR, Goveas SW,
Abraham A. Antioxidant properties of Coscinium fenestratum
stem extracts on Streptozotocin induced type 1 diabetic rats. J
Appl Pharm Sci 2014; 4(1): 29-32.
[7] Yang H, Jin X, Kei Lam CW, Yan SK. Review: oxidative stress
and diabetes mellitus. Clin Chem Lab Med 2011; 49(11): 1773-82.
[8] Gomathi D, Ravikumar G, Kalaiselvi M, Devaki K, Uma C. Ef-
ficacy of Evolvulus alsinoides (L.) L. on insulin and
antioxidants activity in pancreas of streptozotocin induced
diabetic rats. J Diabetes Metab Disord 2013;
http://dx.doi.org/10.1186/2251-6581-12-39.
[9] Sushruta K, Satyanarayana S, Srinivas N, Sekhar JR.
Evaluation of the blood-glucose reducing effects of aqueous
extracts of the selected umbelliferous fruits used in culinary
practices. Trop J Pharm Res 2006; 5(2): 613-7.
[10] Rahmatullah M, Ferdausi D, Mollik AH, Jahan R, Chowdhury
MH, Haque WM. A survey of medicinal plants used by
Kavirajes of Chalna area, Khulna district, Bangladesh. Afr J
Tradit Complement Altern Med 2010; 7(2): 91-7.
[11] Ali H, Houghton PJ, Soumyanath A. alpha-Amylase inhibitory
activity of some Malaysian plants used to treat diabetes; with
particular reference to Phyllanthus amarus. J Ethnopharmacol
2006; 107(3): 449-55.
[12] Bayramoglu G, Senturk H, Bayramoglu A, Uyanoglu M, Colak S,
Ozmen A, et al. Carvacrol partially reverses symptoms of diabetes
in STZ-induced diabetic rats. Cytotechnology 2014; 66(2): 251-7.
[13] Zanin JL, de Carvalho BA, Martineli PS, dos Santos MH, Lago
JH, Sartorelli P, et al. The genus Caesalpinia L.
(Caesalpiniaceae): phytochemical and pharmacological
characteristics. Molecules 2012; 17(7): 7887-902.
[14] Khan HU, Ali I, Khan AU, Naz R, Gilani AH. Antibacterial,
antifungal, antispasmodic and Ca++ antagonist effects of Cae-
salpinia bonducella. Nat Prod Res 2011; 25(4): 444-9.
[15] Adhyapak S, Dighe V. Antidiabetic activity of Caesalpinia bon-
ducella Linn. and Coccinia indica Wight & Arn. in alloxan
induced diabetic rats. Int J Res Pharm Biomed Sci 2013; 4(4):
1287-90.
[16] Kayano AC, Lopes SC, Bueno FG, Cabral EC, Souza-Neiras
WC, Yamauchi LM, et al. In vitro and in vivo assessment of the
anti-malarial activity of. Caesalpinia pluviosa. Malar J 2011;
http:// dx.doi.org/10.1186/1475-2875-10-112.
[17] Wu SQ, Otero M, Unger FM, Goldring MB, Phrutivorapongkul
A, Chiari C, et al. Anti-inflammatory activity of an ethanolic
Cae-salpinia sappan extract in human chondrocytes and
macrophages. J Ethnopharmacol 2011; 138(2): 364-72.
[18] Lorenzi H. Brazilian trees: a guide to the identification and
cultivation of Brazilian native trees. 4th ed. Nova Odessa:
Instituto Plantarum de Estudos da Flora; 2002.
[19] Vasconcelos CF, Maranhão HM, Batista TM, Carneiro EM,
Ferreira F, Costa J, et al. Hypoglycaemic activity and
molecular mechanisms of Caesalpinia ferrea Martius bark
extract on streptozotocin-induced diabetes in Wistar rats. J
Ethnopharmacol 2011; 137(3): 1533-41. ´
[20] Gallão MI, Normando LO, Vieira IGP, Mendes FNP, Ricardo
NM, Brito ES. Morphological, chemical and rheological
properties of the main seed polysaccharide from Caesalpinia
ferrea Mart. Ind Crops Prod 2013; 47: 58-62.
Please cite this article in press as: Hassan SK, et al., Hypoglycemic and antioxidant activities of Caesalpinia ferrea Martius leaf extract in streptozotocin-induced diabetic rats, Asian Pac J
Trop Biomed (2015), http://dx.doi.org/10.1016/j.apjtb.2015.03.004
9
[21] Dias AMA, Rey-Rico A, Oliveira RA, Marceneiro S, Alvarez- Lorenzo C,
Concheiro A, et al. Wound dressings loaded with an anti-inflammatory
juca´ (Libidibia ferrea ) extract using supercritical carbon dioxide
technology. J Supercrit Fluids 2013; 74: 34-45.
[22] Lima SMA, Araujo´ LCC, Sitonioˆ MM, Freitas ACC, Moura SL,
Correia MTS, et al. Anti-inflammatory and analgesic potential of
Caesalpinia ferrea. Rev Bras Farmacogn 2012; 22(1): 169-75.
[23] Sampaio FC, Pereira Mdo S, Dias CS, Costa VC, Conde NC,
Buzalaf MA. In vitro antimicrobial activity of Caesalpinia ferrea
Martius fruits against oral pathogens. J Ethnopharmacol 2009;
124(2): 289-94.
[24] Menezes IA, Moreira IJ, Carvalho AA, Antoniolli AR, Santos
MR. Cardiovascular effects of the aqueous extract from
Caesalpinia ferrea: involvement of ATP-sensitive potassium
channels. Vasc Pharmacol 2007; 47(1): 41-7.
[25] Sousa CC, Gomes SO, Lopes AC, Gomes RL, Britto FB, Lima
PS, et al. Comparison of methods to isolate DNA from
Caesalpinia ferrea. Genet Mol Res 2014; 13(2): 4486-93.
[26] Nozaki H, Hayashi K, Kido M, Kakumoto K, Ikeda S, Matsuura
N, et al. Pauferrol A, a novel chalcone trimer with a
cyclobutane ring from Caesalpinia ferrea mart exhibiting DNA
topoisomerase II inhibition and apoptosis-inducing activity.
Tetrahedron Lett 2007; 48(47): 8290-2.
[27] Ohira S, Takaya K, Mitsui T, Kido M, Kakumoto K, Hayashi K,
et al. New chalcone dimers (I) from Caesalpinia ferrea Mart act
as potent inhibitors of DNA topoisomerase II. Tetrahedron Lett
2013; 54(37): 5052-5.
[28] Nawwar M, El-Mousallami A, Hussein S, Hashem A, Mousa M,
Lindequist U, et al. Three new di-O-glycosyl-C-glucosyl flavones
from the leaves of Caesalpinia ferrea Mart. Z Naturforsch C 2014;
69(9–10): 357-62.
[29] Dawang ND, Datup A. Screening of five medicinal plants for
treatment of typhoid fever and gastroenteritis in central Nigeria.
Glob Eng Technol Rev 2012; 2(9): 1-5.
[30] Mythili T, Ravindhran R. Phytochemical screening and antimi-
crobial activity of Sesbania sesban (L.) Merr. Asian J Pharm Clin
Res 2012; 5(4): 179-82.
[31] Kedare SB, Singh RP. Genesis and development of DPPH method
of antioxidant assay. J Food Sci Technol 2011; 48(4): 412-22.
[32] Chinedu E, Arome D, Ameh FS. A new method for determining
acute toxicity in animal models. Toxicol Int 2013; 20(3): 224-6.
[33] Friedewald WT, Levy RI, Fredrickson DS. Estimation of the
concentration of low-density lipoprotein cholesterol in plasma,
without use of the preparative ultracentrifuge. Clin Chem 1972;
18(6): 499-502.
[34] Necheles TF, Boles TA, Allen DM. Erythrocyte glutathione-
peroxidase deficiency and hemolytic disease of the newborn
in-fant. J Pediatr 1968; 72(3): 319-24.
[35] Minami M, Yoshikawa H. A simplified assay method of super-
oxide dismutase activity for clinical use. Clin Chim Acta 1979;
92(3): 337-42.
[36] Aebi H. Catalase in vitro. Methods Enzymol 1984; 105: 121-6.
[37] Lefevre` G, Beljean-Leymarie M, Beyerle F, Bonnefont-
Rousselot D, Cristol JP, Therond´ P, et al. Evaluation of lipid
peroxidation by measuring thiobarbituric acid reactive
substances. Ann Biol Clin Paris 1998; 56(3): 305-19. French.
[38] Beutler E, Duron O, Kelly BM. Improved method for the deter-
mination of blood glutathione. J Lab Clin Med 1963; 61: 882-8.
[39] Wild S, Roglic G, Green A, Sicree R, King H. Global
prevalence of diabetes: estimates for the year 2000 and
projections for 2030. Diabetes Care 2004; 27(5): 1047-53.
[40] Narvaez´-Mastache JM, Soto C, Delgado G. Hypoglycemic and
antioxidant effects of subcoriacin in normal and streptozotocin-
induced diabetic rats. J Mex Chem Soc 2010; 54(4): 240-4.
[41] Wang R, Ding G, Liang W, Chen C, Yang H. Role of LOX-1
and ROS in oxidized low-density lipoprotein induced epithelial-
mesenchymal transition of NRK52E. Lipids Health Dis 2010; 9:
120.
[42] Xiang FL, Lu X, Strutt B, Hill DJ, Feng Q. NOX2 de ficiency
protects against streptozotocin-induced beta-cell destruction and
development of diabetes in mice. Diabetes 2010; 59(10): 2603-11.
10 Sherien Kamal Hassan et al./Asian Pac J Trop Biomed 2015; ▪(▪): 1–10
[43] Choudhary M, Aggarwal N, Choudhary N, Gupta P, Budhwar
V. Effect of aqueous and alcoholic extract of Sesbania sesban
(Linn.) Merr. root on glycemic control in streptozotocin-induced
diabetic mice. Drug Dev Ther 2014; 5(2): 115-22.
[44] Musabayane CT, Mahlalela N, Shode FO, Ojewole JA. Effects
of Syzygium cordatum (Hochst.) [Myrtaceae] leaf extract on
plasma glucose and hepatic glycogen in streptozotocin-
induced diabetic rats. J Ethnopharmacol 2005; 97(3): 485-90.
[45] Montano ME, Molpeceres V, Mauriz JL, Garzo E, Cruz IB,
Gonzalez´ P, et al. Effect of melatonin supplementation on
food and water intake in streptozotocin-diabetic and non-
diabetic male Wistar rats. Nutr Hosp 2010; 25(6): 931-8.
[46] Juarez´-Rojop IE, D´ıaz-Zagoya JC, Ble-Castillo JL, Miranda-
Osorio PH, Castell-Rodr´ıguez AE, Tovilla-Zarate´ CA, et al. Hy-
poglycemic effect of Carica papaya leaves in streptozotocin-
induced diabetic rats. BMC Complement Altern Med 2012; 12: 236.
[47] Saravanan G, Ponmurugan P, Kumar GPS, Rajarajan T. Antidiabetic
properties of S-allyl cysteine, a garlic component on streptozotocin-
induced diabetes in rats. J Appl Biomed 2009; 7: 151-9.
[48] Uddin N, Hasan MR, Hossain MM, Sarker A, Hasan AH, Islam
AF, et al. In vitro A-amylase inhibitory activity and in vivo
hypoglycemic effect of methanol extract of Citrus macroptera
Montr. fruit. Asian Pac J Trop Biomed 2014; 4(6): 473-9.
[49] Adaramoye OA. Antidiabetic effect of kolaviron, a biflavonoid
complex isolated from Garcinia kola seeds, in Wistar rats. Afr
Health Sci 2012; 12(4): 498-506.
[50] Tundis R, Loizzo MR, Menichini F. Natural products as alpha-
amylase and alpha-glucosidase inhibitors and their hypo-
glycaemic potential in the treatment of diabetes: an update.
Mini Rev Med Chem 2010; 10(4): 315-31.
[51] Girija K, Lakshman K, Udaya C, Sabhya SG, Divya T. Anti-diabetic
and anti-cholesterolemic activity of methanol extracts of three spe-
cies of Amaranthus. Asian Pac J Trop Biomed 2011; 1(2): 133-8.
[52] Singh S, Garg V, Yadav D. Antihyperglycemic and antioxidative
ability of Stevia rebaudiana (Bertoni) leaves in diabetes induced
mice. Int J Pharm Pharm Sci 2013; 5(Suppl. 2): 297-302.
[53] Karim MN, Ahmed KR, Bukht MS, Akter J, Chowdhury HA,
Hossain S, et al. Pattern and predictors of dyslipidemia in
patients with type 2 diabetes mellitus. Diabetes Metab Syndr
2013; 7(2): 95-100.
[54] Pepato MT, Mori DM, Baviera AM, Harami JB, Vendramini RC,
Brunetti IL. Fruit of the jambolan tree ( Eugenia jambolana Lam.) and
experimental diabetes. J Ethnopharmacol 2005; 96(1–2): 43-8.
Please cite this article in press as: Hassan SK, et al., Hypoglycemic and antioxidant activities of Caesalpinia ferrea Martius leaf extract in streptozotocin-induced diabetic rats, Asian Pac J
Trop Biomed (2015), http://dx.doi.org/10.1016/j.apjtb.2015.03.004
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[55] Sharma B, Balomajumder C, Roy P. Hypoglycemic and hypo-
lipidemic effects of flavonoid rich extract from Eugenia
jambolana seeds on streptozotocin induced diabetic rats. Food
Chem Toxicol 2008; 46(7): 2376-83.
[56] Moulisha B, Karan TK, Kar B, Bhattacharya S, Ghosh AK,
Kumar RB, et al. Hepatoprotective activity of Terminalia arjuna
leaf against paracetamol-induced liver damage in rats. Asian J
Chem 2011; 23: 1739-42.
[57] Chitra V, Varma PV, Raju MK, Prakash KJ. Study of antidiabetic
and free radical scavenging activity of the seed extract of Strychnos
nuxvomica. Int J Pharm Pharm Sci 2010; 2(Suppl. 1): 106-10.
[58] Sellamuthu PS, Arulselvan P, Kamalraj S, Fakurazi S, Kandasamy
M. Protective nature of mangiferin on oxidative stress and
antioxidant status in tissues of streptozotocin-induced diabetic
rats. ISRN Phar-macol 2013; http://dx.doi.org/10.1155/2013/750109.
[59] Al-Shiekh AAM, Al-Shati AA, Sarhan MAA. Effect of white tea
extract on antioxidant enzyme activities of streptozotocin-
induced diabetic rats. Egypt Acad J Biol Sci 2014; 6(2): 17-30.
[60] Saravanan G, Ponmurugan P. S-allylcysteine improves
streptozotocin-induced alterations of blood glucose, liver cyto-
chrome P450 2E1, plasma antioxidant system, and adipocytes
hormones in diabetic rats. Int J Endocrinol Metab 2013; 11(4):
e10927.
[61] Lorenzi M. The polyol pathway as a mechanism for diabetic
reti-nopathy: attractive, elusive, and resilient. Exp Diabetes Res
2007; http://dx.doi.org/10.1155/2007/61038.
[62] Cheng D, Liang B, Li Y. Antihyperglycemic effect of Ginkgo
biloba extract in streptozotocin-induced diabetes in rats.
Biomed Res Int 2013; http://dx.doi.org/10.1155/2013/162724.
[63] Almeida DAT, Braga CP, Novelli ELB, Fernandes A. Evaluation of
lipid profile and oxidative stress in STZ-induced rats treated with
antioxidant vitamin. Braz Arch Biol Technol 2012; 55(4): 527-36.
[64] Diao BZ, Jin WR, Yu XJ. Protective effect of polysaccharides from
Inonotus obliquus on streptozotocin-induced diabetic symptoms and
their potential mechanisms in rats. Evid Based Complement Altern
Med 2014; http://dx.doi.org/10.1155/2014/841496.
[65] Robertson RP, Tanaka Y, Takahashi H, Tran PO, Harmon JS.
Prevention of oxidative stress by adenoviral overexpression of
glutathione-related enzymes in pancreatic islets. Ann N. Y Acad
Sci 2005; 1043: 513-20.